Cellular susceptibility and oxidative stress response to menadione of logarithmic, quiescent, and nonquiescent - BioRisk
←
→
Page content transcription
If your browser does not render page correctly, please read the page content below
A peer-reviewed open-access journal
BioRisk 17: 127–138 (2022)
doi: 10.3897/biorisk.17.77320
https://biorisk.pensoft.net
RESEARCH ARTICLE
BioRisk
Cellular susceptibility and oxidative stress response to
menadione of logarithmic, quiescent, and nonquiescent
Saccharomyces cerevisiae cell populations
Polya Galinova Marinovska1, Teodora Ivanova Todorova2,
Krassimir Plamenov Boyadzhiev2, Emiliya Ivanova Pisareva1,
Anna Atanasova Tomova1, Petya Nikolaeva Parvanova2,
Maria Dimitrova2, Stephka Georgieva Chankova2,
Ventsislava Yankova Petrova1
1 Sofia University “St. Kliment Ohridski”, Faculty of Biology, Department of General and Industrial Micro-
biology, 8 Dragan Tsankov Blvd., 1164 Sofia, Bulgaria 2 Institute of Biodiversity and Ecosystems Research,
Bulgarian Academy of Sciences, Department of Ecosystem Research, Environmental Risk Assessment and Con-
servation Biology, 2 Gagarin Str., 1113 Sofia, Bulgaria
Corresponding author: Teodora Todorova (tedi_todorova@yahoo.com)
Academic editor: Roumiana Metcheva | Received 29 October 2021 | Accepted 20 December 2021 | Published 21 April 2022
Citation: Marinovska PG, Todorova TI, Boyadzhiev KP, Pisareva EI, Tomova AA, Parvanova PN, Dimitrova M,
Chankova SG, Petrova VY (2022) Cellular susceptibility and oxidative stress response to menadione of logarithmic,
quiescent, and nonquiescent Saccharomyces cerevisiae cell populations. In: Chankova S, Peneva V, Metcheva R, Beltcheva
M, Vassilev K, Radeva G, Danova K (Eds) Current trends of ecology. BioRisk 17: 127–138. https://doi.org/10.3897/
biorisk.17.77320
Abstract
The aim of the present study was to compare cellular susceptibility and oxidative stress response of S. cerevisiae
logarithmic (log), quiescent (Q), and non-quiescent (NQ) cell populations to menadione – a well-known in-
ducer of oxidative stress. Three main approaches were used: microbiological – cell survival, molecular – con-
stant field gel electrophoresis for detection of DNA double-strand breaks (DSB), and biochemical – measure-
ment of reactive oxygen species (ROS) levels, oxidized proteins, lipid peroxidation, glutathione, superoxide
dismutase (SOD) and catalase on S. cerevisiae haploid strain BY4741. The doses causing 20% (LD20) and
50% (LD50) lethality were calculated. The effect of menadione as a well-known oxidative stress inducer is
compared in the log, Q, and NQ cells. Survival data reveal that Q cells are the most susceptible to menadione
with LD50 corresponding to 9 µM menadione. On the other hand, dose-dependent DSB induction is found
only in Q cells confirming the results shown above. No effect on DSBs levels is observed in log and NQ cells.
Further, the oxidative stress response of the cell populations is clarified. Results show significantly higher lev-
els of SOD and ROS in Q cells than in log cells after the treatment with 100 µM menadione. On the other
side, higher induction of oxidized proteins, malondialdehyde, and glutathione is observed after menadione
treatment of log cells. Our study provides evidence that Saccharomyces cerevisiae quiescent cells are the most
Copyright Polya Galinova Marinovska et al. This is an open access article distributed under the terms of the Creative Commons Attribution License
(CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.128 Polya Galinova Marinovska et al. / BioRisk 17: 127–138 (2022)
susceptible to the menadione action. It might be suggested that the DNA damaging and genotoxic action of
menadione in Saccharomyces cerevisiae quiescent cells could be related to ROS production.
Keywords
Menadione, quiescence, Saccharomyces cerevisiae, stress response
Introduction
Organisms have developed strategies to trigger a stress response when exposed to environ-
mental challenges in order to restore cellular homeostasis (Tagkopoulos et al. 2008; Mitch-
ell et al. 2009). The cellular stress response is thought to be universal and encompasses
a range of cellular functions, including cell cycle control, repair of damaged proteins,
stabilization and repair of DNA and chromatin, cell membrane repair, and more (Kültz
2005). In nature, cells may exist in a proliferative or non-proliferative state (Gangloff and
Arcangioli 2017; Sun and Gresham 2021). The non-proliferative state includes quiescent
or non-quiescent cells (Sun and Gresham 2021). As most of the cells in human tissues are
non-dividing, quiescence is a major form of life (Gangloff and Arcangioli 2017).
Based on this understanding cellular quiescence is of great importance, especially since
studies performed on quiescent cells are still scarce. Such studies in multicellular organisms
are difficult because of the complexity of the signals that control them. One of the possible
solutions is the application of quiescent yeast cells as it is believed that they function simi-
larly to the mammalian and human cells and share similar mechanisms and the same set of
genes involved in the quiescence (Gangloff and Arcangioli 2017; Daskalova et al. 2021a).
Saccharomyces cerevisiae is a widely used test system for studying oxidative stress and
its related consequences. Results obtained on S. cerevisiae could be easily extrapolated
at mammalian, including human level because of homology in genes and conservative
functions of proteins (Foury 1997; Hartwell 2004; Wright et al. 2014). Thus, the ap-
plication of quiescent cells may provide a suitable platform for studying the effect of
various toxic compounds on mammalian and human cells.
The aim of the present study is to compare cellular susceptibility and oxidative
stress response to menadione of S. cerevisiae logarithmic (log), quiescent (Q), and non-
quiescent (NQ) cell populations.
Materials and methods
Saccharomyces cerevisiae strain BY4741
Saccharomyces cerevisiae BY4741 (MATa; his3Δ1; leu2Δ0; met15Δ0; ura3Δ0) obtained
from the EUROSCARF collection was used in the present work. The growth curve of
Saccharomyces cerevisiae BY4741 on YEPD medium is provided as a Suppl. material 1: Fig.
S1. Yeast cells were grown on a standard yeast extract-peptone-dextrose (YEPD) mediumCellular susceptibility to Menadione of Saccharomyces cerevisiae 129
at 30 °C, 204 rpm for 168 h. Yeast media were prepared as described by Sherman et al.
(2001). The growth curve of the strain is provided as a Suppl. material 1: Fig. S1.
Samples were withdrawn at exponential (24 h) and late stationary phase (168 h). Qui-
escent (G0) and non-quiescent cells were isolated from stationary phase yeast population
(168 h) according to the protocol described by Allen et al. (2006). In brief, yeast biomass
in stationary phase (OD540 = 200 (2×109 cells/ml) was layered on Percoll density gradient
and after centrifugation at 400 g (60 min at 20 °C) two layers of cell fractions were formed
– the denser one composed of Go (Q) cells (lower fraction) and a less dense fraction of NQ
cells (upper fraction). Both fractions were separated and microscopically examined. Go cells
were characteristically rounded with thickened cell walls, and no budding cells were ob-
served – these morphological features are typical for the cells in Go state. For comparison,
the stationary phase cell population of NQ cells (upper fraction) was heterogeneous – both
budding, elliptical cells, and deformed, granular and non-budding cells were observed.
Cell survival
Cell suspensions with concentration 1×107 cells/ml were treated with various concen-
trations of menadione (2-methyl-l,4-naphthoquinone, synthetic form of vitamin K)
in the range 1–200 µM for 60 min at 30 °C, 200 rpm. Cells were then centrifuged
(825 g), the supernatant was removed and the pellet was resuspended in a liquid YEPD
medium. Cells were plated on a solid YEPD medium and incubated at 30 °C for 3 days
to evaluate the survival. Doses of lethality (LD20, and LD50) were calculated (Lidanski
1988) by the following formulae:
lgLD50=lgA+(lgB-lgA)/((50-A)/(B-A))
lgLD20=lgA+(lgB-lgA)/((20-A)/(B-A)),
where A – the closest smaller than 50 or 20%, respectively, lethality percentage; lgA- lg
of the concentration corresponding to A; B – the closest higher than 50 or 20%, re-
spectively, lethality percentage; lgB- lg of the concentration corresponding to B.
Cell-free extracts
Isolation of cell-free extracts from log, Q, and NQ cells was carried out according to
the procedure described by Daskalova et al. (2021b) and were used for further bio-
chemical analyses.
Constant field gel electrophoresis (CFGE)
CFGE for detection of DNA double-strand breaks (DSBs) was applied as described in
Todorova et al. (2015, 2019). The levels of DSB induced presented as a mean fraction
of DNA released (FDR) from the wells was quantified by measurement of ethidium
bromide fluorescence using Gene Tool Analyser G: Box (Syngene) and calculated as
described in Chankova et al. (2009).130 Polya Galinova Marinovska et al. / BioRisk 17: 127–138 (2022) Biochemical analysis Oxidative stress markers assay The redox state of logarithmic, quiescent, and non-quiescent yeast cells was assessed through measurement of intracellular levels of accumulated ROS (Kostova et al. 2008), levels of car- bonylated proteins (Mesquita et al. 2014), and oxidized lipids (Hodges et al. 1999). Glutathione measurement The measurement of intracellular glutathione was carried out according to the proce- dure of Zhang (2000). Enzymatic analysis Superoxide dismutase (SOD) and catalase (CAT) enzyme activities were determined spectrophotometrically according to Beauchamp and Fridovich (1971) and Aebi (1984), respectively. Protein content Total intracellular protein was determined according to Lowry et al. (1951). As a standard, bovine serum albumin (Sigma St. Louis, MO, USA) was used. Data analysis The experiments were repeated at least three times from independently grown cultures. Data points in all the figures are mean values. Error bars represent standard errors of mean values. Where no error bars are evident, errors were equal to or less than the symbols. All the calculations were done with GraphPad Prism program, version 6.04 (San Diego, USA). The statistical analysis included the application of Student’s t-test and One-way ANOVA followed by Bonferroni’s post hoc test. P
Cellular susceptibility to Menadione of Saccharomyces cerevisiae 131
Figure 1. Cell survival after menadione treatment A effect of 100 µM menadione on log, Q, and NQ
cell populations B effect of menadione in a concentrations’ range of 1–200 µM on log and Q cells. Each
value represents the mean ± SEM (Standard error of the mean) (n = 3).
Table 1. Levels of lethality calculated after menadione treatment.
Cell populations LD20 (µM) LD50 (µM)
Log 35 199
Quiescent 0.65 9
Two levels of lethality were calculated: LD20 and LD50 (Table 1).
Further, the levels of DSB induced were compared. Our results confirmed the
ones obtained for cell survival. Dose-dependent DSB induction is measured only in
quiescent cells (Fig. 2). The DSB levels measured after the treatment with 150 µM
menadione were 1.5-fold higher than the spontaneous ones. No effect on DSBs levels
is observed in log and NQ cells.
Further experiments were focused on studying the potential differences in the suscep-
tibility based on various markers for oxidative stress – reactive oxygen species, oxidized
proteins, malondialdehyde, intracellular glutathione, superoxide dismutase, and catalase.
Concentration of reactive oxygen species (ROS) after menadione treatment
The ROS measured in the three cell populations are presented in Fig. 3A. The lev-
els measured in G0 cells after menadione treatment are significantly higher – around
3-fold than those measured in the controls. There is a statistically significant but bio-
logically insignificant effect on the ROS levels in log cells. This observation is in a good132 Polya Galinova Marinovska et al. / BioRisk 17: 127–138 (2022)
Figure 2. DSBs induced by various concentrations (50–150 µM) of menadione A induction of DSB
presented as normalized FDR B Q cells C log cells D NQ cells.
Figure 3. Comparative analysis of the levels of reactive oxygen species A oxidized proteins B malondi-
aldehyde C and total glutathione D in S. cerevisiae logarithmic (log), quiescent (Q), and non-quiescent
(NQ) cell populations after the treatment with menadione. Each value represents the mean ± SEM
(Standard error of the mean) (n = 3). Significant differences (* p < 0.05; ** p < 0.001) are presented.
correlation with the cell survival and the DSBs induced in Q cells in comparison with
those observed in log cells.
The constitutive levels of ROS, oxidized proteins, and MDA in NQ cells were
significantly higher than those measured in log and Q cells. Treatment with 100 µM
menadione resulted in significant induction of oxidized proteins and glutathione (Fig.
3B, D). Interestingly, the ROS levels measured in NQ cells were lower after the mena-
dione treatment in comparison with the control levels (Fig. 3A).Cellular susceptibility to Menadione of Saccharomyces cerevisiae 133
Concentration of protein carbonyl groups
Data presented in fig. 3B provides information concerning the concentration of pro-
tein carbonyl groups. Comparing the constitutive levels, around 7-fold higher levels
were measured in Q cells in comparison with the log ones. This could be explained as
a result of the cells’ starvation. Although, the highest quantity – 14 µM/mg was deter-
mined in Q cells the induction was only around 2-fold. Higher induction – around
6-fold was measured in the log cells.
Levels of malondialdehyde (MDA)
Concerning the MDA, comparatively equal constitutive levels were observed between
Q and log cells (Fig. 3C). The NQ cells showed significantly higher MDA levels. As
a result of the menadione treatment the most significant induction of MDA (around
2-fold) was measured in log cells (p < 0.001).
Concentration of glutathione
The GSH concentration in untreated Q cells was 3-fold higher than that in log cells.
Interestingly, menadione treatment did not result in a significant induction of GSH
compared to the untreated control. The GSH concentration was only 2-fold higher
(Fig. 3D). At the same time, the treatment with 100 µM menadione resulted in 7-fold
higher GSH levels in log cells.
Antioxidant enzyme (Superoxide dismutase and Catalase) activity after me-
nadione treatment
Concerning the constitutive levels of the antioxidant enzymes superoxide dismutase and
catalase, differences were obtained. The catalase levels were comparable in the three cell
populations, while SOD was lower in Q cells than in the log and NQ cells (Fig. 4A, B).
Significant induction of SOD was observed in Q cells after the application of
menadione (Fig. 4A). No effect was obtained concerning the catalase levels (Fig. 4B).
Figure 4. Comparative analysis of the response to menadione based on the enzymatic antioxidant system
A superoxide dismutase and B catalase presented as units/mg. Each value represents the mean ± SEM
(Standard error of the mean) (n=3). Significant differences (* p < 0.05; ** p < 0.001) are presented.134 Polya Galinova Marinovska et al. / BioRisk 17: 127–138 (2022)
Discussion
Data presented here provide a comparative analysis of the cellular susceptibility and oxida-
tive stress response to menadione of logarithmic, quiescent, and nonquiescent Saccharo-
myces cerevisiae cell populations. Differences in the cellular susceptibility are obtained de-
pending on the endpoint used. Based on cell survival, DSBs induction, ROS, and SOD Q
cells are more susceptible to menadione. On the other side, higher induction of oxidized
proteins, MDA, and glutathione is observed following menadione treatment of log cells.
The measured increased ROS levels in Q cells correspond well with the decrease
in cell survival and the well-expressed DSB induction. The cytotoxic mechanism of
action of Menadione in G0 cells is stronger, probably due to lower metabolic activity
and higher oxygen levels in the cells. This is in accordance with the report by Fabrizio
and Longo (2008) that quiescent cells are characterized with lower energy consump-
tion and ADP content, which may lead to increased intracellular oxygen levels and
single-electron oxygen reducers. Such conditions may occur during the chronological
aging of yeast cells. On the other hand, the decrease in ROS levels measured after the
treatment with menadione of NQ cells could be explained by the higher percentage of
cells in a terminal state and entering apoptosis (Davidson et al. 2011).
It is already reported that the toxicity of quinones including menadione in S. cer-
evisiae depends on the oxygen presence (Rodrigues-Pousada et al. 2004). This could be
explained by their possible role as catalyzers in the ROS generation via redox-cycling
activity. The cellular response to menadione has been shown to be associated with the
induced synthesis of a large number of proteins, some of which are specific and are
synthesized only upon exposure to this toxic agent (Flattery-O’Brien et al. 1993).
In the present work, log cells showed increased levels of oxidized proteins, MDA,
and glutathione. This could be explained by their increased metabolic activity and a
higher rate of protein synthesis (Daskalova et al. 2021b). Stress-induced toxic oxygen
species, such as superoxide and hydroxyl radicals, damage biological membranes and
other cellular macromolecules, leading to mutations, cancer, or cell death. A direct in-
dicator of the onset of these processes is the appearance of carbonyl groups in proteins,
as well as lipid peroxidation. In addition, the formation of ROS is inevitable under
aerobic conditions due to the reactive nature of molecular oxygen. The action of these
factors individually or jointly can lead to the appearance of oxidative stress – acute or
chronic (Petrova and Kujumdzieva 2010). Oxidative processes that take place during
oxidative stress may lead to reversible or irreversible functional changes in proteins,
which are the main reason for cellular dysfunction. Protein changes are associated with
the formation of carbonyl groups in them. Biochemical analyses have shown that car-
bonyl groups introduced into the side chains of specific amino acids in the active site of
enzymes trigger the initial steps in the degradation of these proteins (von Herrath and
Holzer 1985; Levine and Munro 2002; Grimsrud et al. 2008; Apoorva et al. 2020).
Lipid oxidation occurs through the interaction of ROS with fatty acids in the
membrane lipid layer. This changes the functionality and permeability of biological
membranes and also leads to other disorders. Cell death can be caused by the releaseCellular susceptibility to Menadione of Saccharomyces cerevisiae 135
of cell contents as a result of these changes. Malonaldehyde is the end product of lipid
oxidation. It accumulates in cells and is a highly reactive and toxic electrophilic com-
pound that can form covalently bound products with different proteins. Its concentra-
tion in the cell is used as a biomarker to account for the influence of stress agents. In
our work, the MDA levels remained similar in control and treated Q cells. One of the
explanations could be the thicker cell wall (Daskalova et al. 2021b).
Glutathione plays an important role in protecting the cell against oxidative stress by
protecting it from the toxic effects of ROS through its involvement in mechanisms for
detoxification and regeneration of important cellular antioxidants (Valko et al. 2006).
The antioxidant function of this tripeptide is directly related to the reduction state of
the oxidized GSSG / reduced GSH glutathione pair. More than a few dozen genes have
been identified whose transcription is affected by redox balance in the cell (Allen and
Tresini 2000). It has been found that the GSH: GSSG ratio is of major importance for
this regulation. The glutathione system serves as a cellular redox buffer and changes in
GSH: GSSG balance can lead to oxidation of redox-sensitive cysteine residues in various
proteins (Rahman 2005). Therefore, the increase in intracellular glutathione content
may be one of the adaptive mechanisms to stress in the yeast S. cerevisiae. Glutathione
is a compound with antioxidant and antielectrophilic activity, which suggests its role in
the resistance of cells in a medium with menadione. The accumulation of oxidized glu-
tathione in the cell is an important parameter for measuring the level of oxidative stress.
All enzymes in glutathione metabolism work in an integrated way, allowing the
cell to adapt to different stress conditions (Hayes and Pulford 1995), with de novo
glutathione synthesis being the most important mechanism for increasing levels of
reduced GSH in response to oxidative stress (Rahman 2005). However, the oxidized/
reduced glutathione pair (GSSG/GSH) ratio before and after treatment with mena-
dione remained relatively constant in G0 cells. Controlled changes in GSSG / GSH
contribute to the maintenance of cellular redox potential, which determines resistance
to toxic effects. The stable GSSG / GSH ratio also indicates that in cells of S. cerevisiae
BY4741 strain, menadione exhibits its toxicity through its redox-cyclic mechanism of
action associated with the generation of reactive oxygen species rather than by interac-
tion with reduced glutathione in the cell. In the second case, this would lead to the for-
mation of menadione – S - glutathione conjugates, accompanied by a sharp decrease in
the concentration of intracellular glutathione.
Our study provides evidence that Saccharomyces cerevisiae quiescent cells are the
most susceptible to the menadione action. It might be suggested that the DNA damag-
ing and genotoxic action of menadione in Saccharomyces cerevisiae quiescent cells could
be related to ROS production.
Acknowledgements
This work was supported by a grant from the National Science Fund, Ministry of Edu-
cation and Science, Project No. DH11/10.136 Polya Galinova Marinovska et al. / BioRisk 17: 127–138 (2022)
References
Aebi H (1984) Catalase in Vitro. Methods in Enzymology 105: 121–126. https://doi.
org/10.1016/S0076-6879(84)05016-3
Allen RG, Tresini M (2000) Oxidative stress and gene regulation. Free Radical Biology &
Medicine 28(3): 463–499. https://doi.org/10.1016/S0891-5849(99)00242-7
Allen C, Büttner S, Aragon AD, Thomas JA, Meirelles O, Jaetao JE, Benn D, Ruby SW, Veen-
huis M, Madeo F, Werner-Washburne M (2006) Isolation of quiescent and nonquiescent
cells from yeast stationary-phase cultures. The Journal of Cell Biology 174(1): 89–100.
https://doi.org/10.1083/jcb.200604072
Apoorva S, Behera P, Sajjanar B, Mahawar M (2020) Identification of oxidant susceptible pro-
teins in Salmonella Typhimurium. Molecular Biology Reports 47(3): 2231–2242. https://
doi.org/10.1007/s11033-020-05328-3
Beauchamp C, Fridovich I (1971) Superoxide Dismutase: Improved Assays and an Assay
Applicable to Acrylamide Gels. Analytical Biochemistry 44(1): 276–287. https://doi.
org/10.1016/0003-2697(71)90370-8
Chankova SG, Yurina NP, Dimova EG, Ermohina OV, Oleskina YP, Dimitrova MT, Bryant
PE (2009) Pretreatment with heat does not affect double-strand breaks DNA rejoining
in Chlamydomonas reinhardtii. Journal of Thermal Biology 34(7): 332–336. https://doi.
org/10.1016/j.jtherbio.2009.06.002
Daskalova AV, Tomova AA, Kujumdzieva AV, Velkova LG, Dolashka PA, Petrova VY (2021a)
Menadione and hydrogen peroxide trigger specific alterations in RNA polymerases pro-
files in quiescent Saccharomyces cerevisiae cells. Biotechnology, Biotechnological Equipment
35(1): 1190–1199. https://doi.org/10.1080/13102818.2021.1941255
Daskalova A, Petrova V, Velkova L, Kujumdzieva A, Tomova A, Voelter W, Dolashka P (2021b)
Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and pro-
liferating state before and after toxic stress. Biotechnology, Biotechnological Equipment
35(1): 366–376. https://doi.org/10.1080/13102818.2021.1879677
Davidson GS, Joe RM, Roy S, Meirelles O, Allen CP, Wilson MR, Tapia PH, Manzanilla
EE, Dodson AE, Chakraborty S, Carter M, Young S, Edwards B, Sklar L, Werner-Wash-
burne M (2011) The proteomics of quiescent and nonquiescent cell differentiation in
yeast stationary-phase cultures. Molecular Biology of the Cell 22(7): 988–998. https://doi.
org/10.1091/mbc.e10-06-0499
Fabrizio P, Longo VD (2008) Chronological aging-induced apoptosis in yeast. Biochimica et
Biophysica Acta 1783(7): 1280–1285. https://doi.org/10.1016/j.bbamcr.2008.03.017
Flattery-O‘Brien J, Collinson LP, Dawes IW (1993) Saccharomyces cerevisiae has an induci-
ble response to menadione which differs from that to hydrogen peroxide. Microbiology
139(3): 501–507. https://doi.org/10.1099/00221287-139-3-501
Foury F (1997) Human genetic diseases – a cross-talk between man and yeast. Gene 195(1):
1–10. https://doi.org/10.1016/S0378-1119(97)00140-6
Gangloff S, Arcangioli B (2017) DNA repair and mutations during quiescence in yeast. FEMS
Yeast Research 17(1): fox002. https://doi.org/10.1093/femsyr/fox002Cellular susceptibility to Menadione of Saccharomyces cerevisiae 137
Grimsrud PA, Xie H, Griffin TJ, Bernlohr DA (2008) Oxidative stress and covalent modifica-
tion of protein with bioactive aldehydes. The Journal of Biological Chemistry 283(32):
21837–21841. https://doi.org/10.1074/jbc.R700019200
Hartwell LH (2004) Yeast and cancer. Bioscience Reports 24(4–5): 523–544. https://doi.
org/10.1007/s10540-005-2743-6
Hayes JD, Pulford DJ (1995) The glutathione S-transferase supergene family: Regulation of
GST and the contribution of the lsoenzymes to cancer chemoprotection and drug resist-
ance part I. Critical Reviews in Biochemistry and Molecular Biology 30(6): 445–520.
https://doi.org/10.3109/10409239509083491
Hodges PE, McKee AH, Davis BP, Payne WE, Garrels JI (1999) The Yeast Proteome Database
(YPD): A model for the organization and presentation of genome-wide functional data.
Nucleic Acids Research 27(1): 69–73. https://doi.org/10.1093/nar/27.1.69
Kostova I, Traykova M, Rastogi VK (2008) New lanthanide complexes with antioxidant activ-
ity. Medicinal Chemistry (Shariqah, United Arab Emirates) 4(4): 371–378. https://doi.
org/10.2174/157340608784872181
Kültz D (2005) Molecular and evolutionary basis of the cellular stress response. Annual Review of
Physiology 67(1): 225–257. https://doi.org/10.1146/annurev.physiol.67.040403.103635
Levine TP, Munro S (2002) Targeting of Golgi-specific pleckstrin homology domains involves
both PtdIns 4-kinase-dependent and -independent components. Current Biology 12(9):
695–704. https://doi.org/10.1016/S0960-9822(02)00779-0
Lidanski T (1988) Statistical methods in biology and agriculture. Zemizdat, Sofia, 375. [in
Bulgarian]
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin
phenol reagent. Biological Chemistry 193(1): 265–275. https://doi.org/10.1016/S0021-
9258(19)52451-6
Mesquita CS, Oliveira R, Bento F, Geraldo D, Rodrigues JV, Marcos JC (2014) Simplified
2, 4-dinitrophenylhydrazine spectrophotometric assay for quantification of carbonyls
in oxidized proteins. Analytical Biochemistry 458: 69–71. https://doi.org/10.1016/j.
ab.2014.04.034
Mitchell A, Romano GH, Groisman B, Yona A, Dekel E, Kupiec M, Dahan O, Pilpel Y (2009)
Adaptive prediction of environmental changes by microorganisms. Nature 460(7252):
220–224. https://doi.org/10.1038/nature08112
Petrova VY, Kujumdzieva AV (2010) Robustness of Saccharomyces Cerevisiae Genome to An-
tioxidative Stress, Biotechnology & Biotechnological Equipment 24(sup1): 474–483.
https://doi.org/10.1080/13102818.2010.10817886
Rahman I (2005) Regulation of glutathione in inflammation and chronic diseases. Life Sci-
ences 75(1): 93–103. https://doi.org/10.1016/j.mrfmmm.2005.02.025
Rodrigues-Pousada CA, Nevitt T, Menezes R, Azevedo D, Pereira J, Amaral C (2004) Yeast
activator proteins and stress response: An overview. FEBS Letters 567(1): 80–85. https://
doi.org/10.1016/j.febslet.2004.03.119
Sherman F, Fink GR, Hicks GB (2001) Methods in Yeast Genetics: A Laboratory Manual.
Cold Spring Harbor Laboratory Press, Plainview, NY, 2001.138 Polya Galinova Marinovska et al. / BioRisk 17: 127–138 (2022)
Sun S, Gresham D (2021) Cellular quiescence in budding yeast. Yeast (Chichester, England)
38(1): 12–29. https://doi.org/10.1002/yea.3545
Tagkopoulos I, Liu YC, Tavazoie S (2008) Predictive behavior within microbial genetic net-
works. Science 320: https://doi.org/10.1126/science.1154456
Todorova T, Miteva D, Chankova S (2015) DNA damaging effect of zeocin and methyl meth-
anesulfonate in Saccharomyces Cerevisiae measured by CFGE. Dokladi na Bulgarskata Aka-
demiâ na Naukite 68(1).
Todorova T, Miteva D, Chankova S (2019) DNA susceptibility of Saccharomyces cerevisiae to
Zeocin depends on the growth phase. International Microbiology 22(4): 419–428. https://
doi.org/10.1007/s10123-019-00065-5
Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M (2006) Free radicals, metals and anti-
oxidants in oxidative stress-induced cancer. Chemico-Biological Interactions 160(1): 1–40.
https://doi.org/10.1016/j.cbi.2005.12.009
von Herrath M, Holzer H (1985) Oxidative inactivation of yeast fructose-1,6-bisphosphatase.
Progress in Clinical and Biological Research 180: 329–340.
Wright EP, Padula MP, Higgins VJ, Aldrich-Wright JR, Coorssen JR (2014) A Systems Biology
Approach to Understanding the Mechanisms of Action of an Alternative Anticancer Com-
pound in Comparison to Cisplatin. Proteomes 2(4): 501–526. https://doi.org/10.3390/
proteomes2040501
Zhang Y (2000) Role of glutathione in the accumulation of anticarcinogenic isothiocyanates
and their glutathione conjugates by murine hepatoma cells. Carcinogenesis 21(6): 1175–
1182. https://doi.org/10.1093/carcin/21.6.1175
Supplementary material 1
Figure S1
Authors: Polya Galinova Marinovska, Teodora Ivanova Todorova, Krassimir Plamenov
Boyadzhiev, Emiliya Ivanova Pisareva, Anna Atanasova Tomova, Petya Nikolaeva
Parvanova, Maria Dimitrova, Stephka Georgieva Chankova, Ventsislava Yankova Petrova
Data type: jpg file
Explanation note: Fig. S1. Growth curve of Saccharomyces cerevisiae BY4741 and glu-
cose assimilation in batch cultivation on YEPD media at 30 °C, 204 rpm for 168 h.
Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.
Link: https://doi.org/10.3897/biorisk.17.77320.suppl1You can also read
